|
type |
Journal Article |
authors |
Zhao G, Liu J, Liu X, Chen M, Zhang H, Wang PG. |
title |
Cloning and characterization of GDP-perosamine synthetase (Per) from Escherichia coli O157:H7 and synthesis of GDP-perosamine in vitro
|
journal |
Biochem Biophys Res Commun |
Activity |
2.6.1.102 |
Family |
2.6.1.102 |
sel |
selected |
ui |
17888872 |
year |
(2007) |
volume |
363 |
number |
3 |
pages |
525-30 |
| |
abstract |
GDP-perosamine synthetase (Per, E.C. not yet classified) is important to the synthesis of Escherichia coli O157:H7 O-antigen. The mutant in per gene can disrupt the synthesis of O157 O-antigen. In this study, GDP-perosamine synthetase was cloned from E. coli O157:H7 and over-expressed in E. coli BL21 (DE3). The recombinant His-tagged Per fusion protein was a decamer with molecular weight of 431 kDa. The optimal pH value of this recombinant protein was 7.5. The divalent ions had no significant effect on Per-catalyzed reaction. The K(m) and K(cat)/K(m) for GDP-4-keto-6-deoxy-d-mannose were 0.09 mM and 2.1 x 10(5)M(-1)S(-1), and those for l-glutamate were 2mM and 0.52 x 10(5)M(-1)S(-1), respectively. Per was used to synthesize GDP-perosamine from GDP-mannose together with recombinant GDP-mannose dehydratase (GMD, E.C. 4.2.1.47). The purified GDP-perosamine was identified by MS and NMR. In summary, this work provided a feasible approach for the synthesis of GDP-perosamine which can lead to the study of LPS biosynthesis of pathogenic E. coli O157:H7. |
last changed |
2014/02/20 12:05 |
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